Polyepitopic protein fragments of the e6 and e7 proteins of hpv, their production and their use particularly in vaccination

ABSTRACT

Polyepitopic peptides of E6 and E7 proteins of Human Papillomavirus, their production, and methods of treating pathologies in which a polyepitopic peptide of the E6 and E7 protein of Human Papillomavirus is recognized by the cellular immune system.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a division of co-pending application Ser. No.10/858,384 filed on Jun. 2, 2004, which is a divisional application U.S.application Ser. No. 09/980,523, filed on Apr. 29, 2002, now U.S. Pat.No. 6,783,763 which was a national phase of PCT InternationalApplication No. PCT/FR00/01513 filed on May 31, 2000 under 35 U.S.C. §371, the entire contents of each application are hereby incorporated byreference.

BACKGROUND OF THE INVENTION

The present invention has for its object polyepitopic protein fragments,such as those of the E6 and E7 proteins of human papillomavirus, or ofthe human p53 protein, their process of production, and their uses,particularly in the field of therapeutic or preventive vaccination.

SUMMARY OF THE INVENTION

The invention more particularly has for its object the use ofpolyepitopic fragments of a predetermined protein for the preparation ofmedications adapted for the prevention or treatment of pathologies inwhich said protein is recognized by the cellular immune system.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows an E6 protein of HPV 16;

FIG. 2 shows an E7 protein of HPV 16.

DETAILED DESCRIPTION OF THE INVENTION

Preferably, said polyepitopic fragments are such that their N-terminalamino acid corresponds to the N-terminal amino acid of the epitopelocated upstream of one or several other epitopes of a polyepitopicregion of said protein, and their C-terminal amino acid corresponds tothe C-terminal amino acid of the epitope located downstream of theabove-mentioned epitope or epitopes of said polyepitopic region.

Thus, the above-mentioned polyepitopic protein fragments of the presentinvention correspond preferably to the polyepitopic regions of apredetermined protein, namely to the regions containing several epitopesrecognized by the T cells in association with the different molecules ofthe major complex of histocompatibility (MCH), said regions beingselected from those having the characteristic of being degraded in vitroin shorter peptides by proteasomes, such as the 20S proteasome, when theprotein fragment tested is placed in the presence of said proteasome,particularly according to the following detailed method. The proteinfragment (about 75 μg when it is a polypeptide of about 30 amino acids)is incubated at 37° C. with about 15 μg of 20S proteasome (CalbiochemRef 539150, La Jolla, Calif., USA) in 500 μl of the following buffer: 20mM Tris-HCl pH8, 0.5 mM EDTA.

Aliquots of 50 μl are removed after incubation times of 24 and 48 hours,and are analyzed by high pressure liquid chromatography (HPLC). Thedigestion products of the proteasomes are separated by RP-HPLC (PerkinElmer) by using a C18 column and an acetonitrile gradient (from 0 to100% containing 0.1% trifluoroacetic acid, for 90 minutes, elution rate0.8 ml/min). The cleavage products are detected at 214 nm by anabsorption detector (759A, Applied Biosystems).

Preferably, the polyepitopic regions defined above have thecharacteristic of containing hydrophobic amino acids.

The different epitopes of the polyepitopic region of the predeterminedprotein, and delimiting the epitopic protein fragments, are preferablyselected from the peptides;

-   -   binding to a predetermined molecule of MCH, particularly to a        molecule of the predetermined HLA type, and this from        concentrations of about 10⁻⁶ M to about 10⁻¹⁰ M in peptide for        concentrations of about 10⁻⁷ M of HLA molecules, particularly        under the conditions described hereafter,    -   and forming a stable complex with said MCH molecule, namely        particularly a complex in which said peptide remains bound to        said molecule for at least about 3 hours at 37° C.

By way of illustration, the above-mentioned epitopes of the inventionare selected from among the peptides adapted:

-   -   on the one hand of associating with the molecules of MCH,        particularly by using the following method:        -   incubation (particularly for about 2 hours at 25° C., then            about 15 hours at 4° C.) of the peptide in the presence of            MCH molecules, from the lysis of human or animal cells, or            purified particularly by affinity chromatography from human            or animal cell lines,        -   trapping complexes formed during the preceding step on a            solid support covered with a first antibody, particularly            monoclonal, recognizing specifically the CMH molecules in            their configuration dependent on their connection to said            peptide,        -   addition to the preceding solid support of a second marked            antibody, particularly by coupling to a radioactive,            enzymatic or fluorescent marker, said marked antibody            recognizing specifically either the long chains of MCH in            their configuration dependent on their connection to the            peptide, or the short chain of MCH or the β2-microglobulin            binding specifically to the different long chains of the MCH            in their above-mentioned configuration,        -   detection, after rinsing of the solid support, of the            possible presence of the second marked antibody remaining            fixed on the solid support, signifying an association effect            between the molecules of MCH and the peptide studied,    -   and, on the other hand, forming a complex with said MCH        molecules, whose stability can be evaluated by the use of a        method of following according to time the connection established        between the peptide and the MCH molecules, this method being        preferably carried out according to a protocol identical to the        preceding method, but in which the incubation step of the        peptide in the presence of MCH molecules on the solid support        covered with said first antibody, is preceded by a preliminary        step of eliminating the free peptide adapted that may be present        in the reaction medium, particularly by washing the solid        support, said incubation step being carried out (preferably at a        temperature of 37° C.) for variable times of 1 hours, 3 hours, 5        hours, 24 hours and 48 hours.

As mentioned above, the epitopes of the invention should be recognizedby the T cells in association with the MCH molecules and associate withthese latter, particularly in the framework of the practice of therecognition test described above. This association can be weak(detectable at concentrations of peptide analogs of the order of 10⁻⁴ to10⁻⁵ M), intermediate (detectable at concentrations of peptide analogsof the order of 10⁻⁶ to 10⁻⁷ M), or strong (detectable at concentrationsof peptide analogs of the order of 10⁻⁸ to 10⁻⁹ M). The peptidesassociated with the MCH molecules in the scope of the present inventionare preferably adapted to bond during at least about 3 hours, to saidMCH molecules.

The invention more particularly has for its object the epitopes (alsodesignated peptides above and hereafter) as described above andcharacterized in that they are selected from among those adapted:

-   -   to induce in vitro cytolysis by cytotoxic T lymphocytes, of        target cells having at their surface the above-mentioned peptide        associated with the MCH molecules, said cytotoxic T lymphocytes        being preferably removed from a patient having a pathology in        which the peptide studied is implied,    -   and inducing in vitro the secretion of cytokines (or        interleukines) by the above-mentioned cytotoxic T lymphocytes,        particularly IL-2, IL-4 or γ interferon.

As the case may be, the above-mentioned epitopes are selected from thoseable to induce in vitro the appearance and the growth of cytotoxic Tlymphocytes from animal or human cells, particularly from peripheralblood mononucleated cells (PBMC), in the presence of factors necessaryfor the growth and differentiation of the cytotoxic T cells.

The polyepitopic protein fragments of the invention are moreovercharacterized in that they are adapted to contain CD4 epitopesrecognized by auxiliary T cells in association with the MCH molecules ofclass II, this property favoring the induction and maintenance of theCD8⁺ T cells recognizing the epitopes comprised in said fragments.

The present invention is illustrated with the help of FIGS. 1 (SEQ IDNO: 2) and 2 (SEQ ID NO: 12), showing respectively peptide sequences ofthe E6 (SEQ ID NO: 2) and E7 (SEQ ID NO: 12) proteins of the strain 16of the human papillomavirus (HPV 16), as well as the polyepitopicfragments of the invention, and the epitopes within these fragments.

The invention more particularly has for its object the polyepitopicfragments of the E6 and E7 protein of HPV, and more particularly thoseof the E6 protein shown in FIG. 1, or by SEQ ID NO: 2, or those of theE7 protein, shown in FIG. 2, or by SEQ ID NO: 11, of HPV 16,characterized in that they comprise a peptide sequence of about 15 toabout 30 amino acids, this peptide sequence containing the amino acidsequences of at least 3 different epitopes, and preferably at least 4different epitopes binding stably to HLA molecules of identical ordifferent type, when these epitopes are obtained by enzymaticdegradation of said peptide sequence, particularly in the proteasome,such that at least 4 HLA molecules of different types, and preferably atleast 5 HLA molecules of different types, bind to these epitopes, these4 or 5 HLA molecules being selected from those of type A1, A2, A3, A11,A24, A29, B7, B8, B18, B27, B35, B44, B51, and B62.

Preferably, the polyepitopic fragments according to the invention aresuch that the number of amino acids of their peptide sequence is greaterthan or equal to 17, and less than or equal to 30.

The invention relates more particularly to the polyepitopic fragments ofthe E6 protein of HPV defined above, characterized in that they comprisea peptide sequence of about 15 to 30 amino acids, this peptide sequencecontaining amino acid sequences of at least 5 different epitopes, andpreferably at least 6 different epitopes binding stably to HLA moleculesof identical or different type, when these epitopes are obtained byenzymatic degradation of said peptide sequence, particularly in theproteasome, such that at least 6 HLA molecules of different types, andpreferably at least 7 HLA molecules of different types, bind to theseepitopes, these 6 or 7 HLA molecules being selected from those of typeA1, A2, A3, A11, A24, A29, B7, B8, B18, B27, B35, B44, and B51.

Preferably, the polyepitopic fragments of the E6 protein according tothe invention are such that the number of amino acids of their peptidesequence is greater than or equal to 20 (preferably greater than orequal to 22), and less than or equal to 30.

Also preferably, the above-mentioned polyepitopic fragments of the E6protein of HPV, are characterized in that they all comprise an epitopebinding to the HLA molecule of type B35, an epitope binding to the HLAmolecule of type B44, and an epitope binding to the HLA molecule of typeB51.

The invention more particularly has for its object the polyepitopicfragment of the E6 protein of HPV as defined above, characterized inthat it corresponds to the fragment of 30 amino acids delimited by theamino acids located in positions 15 and 44 of the peptide sequence ofthe E6 protein of HPV, and characterized by the peptide sequence SEQ IDNO: 4 as follows:

(15)RPRKLPQLCTELQTTIHDIILECVYCKQQL(44)

said fragment containing 9 epitopes binding stably to at least one ofthe 8 HLA molecules of the following types: A2, A11, A29, B7, B8, B35,B44, or B51, said epitopes being the following:

-   -   (15)RPRKLPQL(22) (residues 15 to 22 of SEQ ID NO: 2) binding        stably to HLA molecules of the B7 or B35 type,    -   (18)KLPQLCTEL(26) (residues 18 to 26 of SEQ ID NO: 2) binding        stably to HLA molecules of the A2 type,    -   (19)LPQLCTEL(26) (residues 19 to 26 of SEQ ID NO: 2) binding        stably to HLA molecules of the B51 type,    -   (21)QLCTELQTTI(30) (residues 21 to 30 of SEQ ID NO: 2) binding        stably to HLA molecules of the A2 type,    -   (24)TELQTTIHDI(33) (residues 24 to 33 of SEQ ID NO: 2) binding        stably to HLA molecules of the A29 or B44 type,    -   (29)TIHDIILRCV(38) (residues 29 to 38 of SEQ ID NO: 2)binding        stably to HLA molecules of the A2 type,    -   (33)IILECVYCK(41) (residues 33 to 41 of SEQ ID NO: 2) binding        stably to HLA molecules of the A11 type,    -   (35)LECVYCKQQL(44) (residues 35 to 44 of SEQ ID NO: 2) binding        stably to HLA molecules of the A29 or B44 type,    -   (37)CVYCKQQL(44) (residues 37 to 44 of SEQ ID NO: 2) binding        stably to HLA molecules of the B8 type.

The invention also relates to the polyepitopic fragment of the E6protein of HPV as defined above, characterized in that it corresponds tothe fragment of 17 amino acids delimited by the amino acids located atpositions 46 and 62, or to the fragment of 22 amino acids delimited bythe amino acids located at positions 46 and 67 of the peptide sequenceof the E6 protein of HPV, this latter fragment being characterized bythe peptide sequence SEQ ID NO: 6 as follows:

(46)RREVYDFAFRDLCIVYRDGNPY(67)

said fragment containing 6 epitopes binding stably to at least one ofthe 10 HLA molecules of the following types: A2, A3, A11, A24, A29, B7,B27, B35, B44, or B51, said epitopes being the following:

-   -   (46)RREVYDFAFR(55) (residues 46 to 55 of SEQ ID NO: 2) binding        stably to HLA molecules of the B27 type,    -   (49)VYDFAFRDL(57) (residues 49 to 57 of SEQ ID NO: 2) binding        stably to HLA molecules of the A24 type,    -   (50)YDFAFRDL(57) (residues 50 to 57 of SEQ ID NO: 2) binding        stably to HLA molecules of the A29 or B44 type,    -   (52)FAFRDLCIV(60) (residues 52 to 60 of SEQ ID NO: 2) binding        stably to HLA molecules of the A2, B35, B51, or B7 type,    -   (54)FRDLCIVYR(62) (residues 54 to 62 of SEQ ID NO: 2) binding        stably to HLA molecules of the A3 or A11 type,    -   (59)IVYRDGNPY(67) (residues 59 to 67 of SEQ ID NO: 2) binding        stably to HLA molecules of the A3 or A11 type.

The invention also has for its object the polyepitopic fragment of theE6 protein of HPV as defined above, characterized in that it correspondsto the fragment of 29 amino acids delimited by the amino acids locatedat positions 80 and 108 of the peptide sequence of the E6 protein ofHPV, this latter fragment being characterized by the peptide sequenceSEQ ID NO: 8 as follows:

(8O)ISEYRHYCYSLYGTTLEQQYNKPLCDLLI(108)

said fragment containing 6 epitopes binding stably to at least 10 HLAmolecules of the following types: A1, A3, A11, A24, A29, B7, B18, B35,B44, or B51, said epitopes being the following:

-   -   (80)ISEYRHYCY(88) (residues 80 to 88 of SEQ ID NO: 2) binding        stably to HLA molecules of the A1 or B18 type,    -   (81)SEYRHYCY(88) (residues 81 to 88 of SEQ ID NO: 2) binding        stably to HLA molecules of the A29 or B44 type,    -   (87)CYSLYGTTL(95) (residues 87 to 95 of SEQ ID NO: 2) binding        stably to HLA molecules of the A24 type,    -   (94)TLEQQYNK101) (residues 94 to 101 of SEQ ID NO: 2)binding        stably to HLA molecules of the A3 or A11 type,    -   (95)LEQQYNKPL(103) (residues 95 to 103 of SEQ ID NO: 2) binding        stably to HLA molecules of the A29 or B44 type,

(101)KPLCDLLI(108) (residues 101 to 108 of SEQ ID NO: 2) binding stablyto HLA molecules of the B7, B35 or B51 type.

The invention more particularly has for its object the polyepitopicfragment of the E6 protein of HPV as defined above, characterized inthat it corresponds to the fragment of 22 amino acids delimited by theamino acids located at positions 118 and 139 of the peptide sequence ofthe E6 protein of HPV, this latter fragment being characterized by thepeptide sequence SEQ ID NO: 10 as follows:

(118)CPEEKQRHLDKKQRFHNIRGRW(139)

said fragment containing 6 epitopes binding stably to at least one ofthe 7 HLA molecules of the following types: A24, B8, B18, B27, B35, B44,or B51, said epitopes being the following:

-   -   (118)CPEEKQRHL(126) (residues 118 to 126 of SEQ ID NO: 2)        binding stably to HLA molecules of the B8, B18, B35, B51 type,    -   (119)PEEKQRHL(126) (residues 119 to 126 of SEQ ID NO: 2) binding        stably to HLA molecules of the B44 type,    -   (127)DKKQRFHNI(135) (residues 127 to 135 of SEQ ID NO: 2)        binding stably to HLA molecules of the B8 type,    -   (128)KKQRFHNIR(136) (residues 128 to 136 of SEQ ID NO: 2)        binding stably to HLA molecules of the B27 type,    -   (130)QRFHNIRGRW(139) (residues 130 to 139 of SEQ ID NO: 2)        binding stably to HLA molecules of the B27 type,    -   (131)RFHNIRGRW(139) (residues 131 to 139 of SEQ ID NO: 2)        binding stably to HLA molecules of the A24 type.

The invention also relates to the polyepitopic fragments of the E7protein of HPV as defined above, characterized in that they comprise apeptide sequence of about 15 to 30 amino acids, this peptide sequencecontaining the amino acid sequences of at least 3 different epitopes,and preferably of at least 4 different epitopes binding stably to HLAmolecules of the identical or different type, when these epitopes areobtained by enzymatic degradation of said peptide sequence, particularlyin the proteasome, such that at least 4 HLA molecules of differenttypes, and preferably at least 5 HLA molecules of different types bindto these epitopes, these 4 or 5 HLA molecules being selected from thoseof type A1, A2, A3, A11, A29, B7, B18, B35, B44, and B62.

Preferably, the polyepitopic fragments of the E7 protein according tothe invention are such that the number of amino acids of the peptidesequence is greater than or equal to 17, and less than or equal to 23.

Again preferably, the polyepitopic fragments of the E7 protein of theabove-mentioned HPV, are characterized in that they all comprise anepitope binding to the HLA molecule of type B44.

The invention more particularly has for its object the polyepitopicfragment of the E7 protein of HPV as defined above, characterized inthat it corresponds to the fragment of 23 amino acids delimited by theamino acids located in positions 3 and 25 of the peptide sequence of theE7 protein of HPV, this latter fragment being characterized by thepeptide sequence SEQ ID NO: 14 as follows:

(3)GDTPTLHEYMLDLQPETTDLYCY(25)

said fragment containing 5 epitopes binding stably to at least one ofthe 6 HLA molecules of the following type: A1, A2, B18, B35, B44 or B62,said epitopes being the following:

-   -   (3)GDTPTLHEY(11) (residues 3 to 11 of SEQ ID NO: 12) binding        stably to HLA molecules of the B44 type,    -   (5)TPTLHEYML(13) (residues 5 to 13 of SEQ ID NO: 12) binding        stably to HLA molecules of the B35 type,    -   (11)YMLDLQPETT(20) (residues 11 to 20 of SEQ ID NO: 12) binding        stably to HLA molecules of the A2 type,    -   (15)LQPETTDLY(23) (residues 15 to 23 of SEQ ID NO: 12) binding        stably to HLA molecules of the B62 type,    -   (16)QPETTDLYCY(25) (residues 16 to 25 of SEQ ID NO: 12) binding        stably to HLA molecules of the A1 or B18 type.

The invention also relates to the polyepitopic fragment of the E7protein of HPV as defined above, characterized in that it corresponds tothe fragment of 17 amino acids delimited by the amino acids located inpositions 44 and 60 of the peptide sequence of the E7 protein of HPV,this latter fragment being characterized by the peptide sequence SEQ IDNO: 16 as follows:

(44)QAEPDRAHYNIVTFCCK(60)

said fragment containing 4 epitopes binding stably to at least one ofthe 6 HLA molecules of the following types: A1, A3, A11, A29, B7, B18,B35, or B44, said epitopes being the following:

-   -   (44)QAEPDRAHY(52) (residues 44 to 52 of SEQ ID NO: 12) binding        stably to HLA molecules of the A1 or B18 type,    -   (45)AEPDRAHY(52) (residues 45 to 52 of SEQ ID NO: 12) binding        stably to HLA molecules of the A29 or B44 type,    -   (46)EPDRAHYNIV(55) (residues 46 to 55 of SEQ ID NO: 12) binding        stably to HLA molecules of the B7 or B35 type,    -   (53)NIVTFCCK(60) (residues 53 to 60 of SEQ ID NO: 12) binding        stably to HLA molecules of the A3 or A11 type.

The invention also has for its object the polyepitopic fragment of theE7 protein of HPV as defined above, characterized in that it correspondsto the fragment of 19 amino acids delimited by the amino acids locatedin positions 79 and 97 of the peptide sequence of the E7 protein of HPV,this latter fragment being characterized by the peptide sequence SEQ IDNO: 18 as follows:

(79)LEDLLMGTLGIVCPICSQK(97)

said fragment containing 4 epitopes binding stably to at least one ofthe 5 HLA molecules of the following types: A2, A3, A11, A29 or B44,said epitopes being the following:

-   -   (79)LEDLLMGTL(87) (residues 79 to 87 of SEQ ID NO: 12) binding        stably to HLA molecules of the A29 or B44 type,    -   (82)LLMGTLGIV(90) (residues 82 to 90 of SEQ ID NO: 12) binding        stably to HLA molecules of the A2 type,    -   (86)TLGIVCPI(93) (residues 86 to 93 of SEQ ID NO: 12) binding        stably to HLA molecules of the A2 type,    -   (89)IVCPICSQK(97) (residues 89 to 97 of SEQ ID NO: 12) binding        stably to HLA molecules of the A3 or A11 type.

The invention also has for its object the polyepitopic fragments of thep53 human protein characterized in that they comprise a peptide sequenceof about 20 to about 35 amino acids, this latter containing amino acidsequences of at least three different epitopes binding stably to HLAmolecules of identical or different type, when these epitopes areobtained by enzymatic degradation of said peptide sequence, particularlyin the proteasome, such that at least 3 HLA molecules of different typeswill be recognized by said epitopes and will bind to these latter, these3 HLA molecules being selected from those of type A1, A2, A3, A24, B7,B8, B27, B35, B44 and B62.

The invention also relates to the polyepitopic fragments of the p53human protein mentioned above, characterized in that they comprise apeptide sequence of about 20 to about 35 amino acids, this lattercontaining the amino acid sequences of at least 5 different epitopes,and preferably of at least 6 different epitopes binding to HLA moleculesof identical or different type, such that at least 3 HLA molecules ofdifferent types, and preferably at least 4 HLA molecules of differenttypes will be recognized by said epitopes and will bind to these latter,these 3 or 4 HLA molecules being selected from those of type A2, A24,B27, B35, B44 and B62.

The invention more particularly has for its object the polyepitopicfragment of the p53 human protein as defined above, characterized inthat it corresponds to the fragment of 32 amino acids delimited by theamino acids located in positions 106 and 137 of the peptide sequence ofthe p53 protein, or to the fragment of 36 amino acids delimited by theamino acids in positions 102 and 137 of said peptide sequence, thislatter fragment being characterized by the following peptide sequence:

(SEQ ID NO: 19) (102)TYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQL(137)

said fragment containing 6 epitopes binding stably to at least one ofthe 4 HLA molecules of the following types: A2, A24, B35 or B62, saidepitopes being the following:

-   -   (102)TYQGSYGFRL(111) (residues 1 to 10 of SEQ ID NO: 19) binding        stably to HLA molecules of the A24 type,    -   (105)GSYGFRLGFL(114) (residues 4 to 13 of SEQ ID NO: 19) binding        stably to HLA molecules of the B35 type,    -   (106)SYGFRLGFL(114) (residues 5 to 13 of SEQ ID NO: 19) binding        stably to HLA molecules of the A24 type,    -   (118)TAKSVTCTY(126) (residues 17 to 25 of SEQ ID NO: 19) binding        stably to HLA molecules of the B62 type,    -   (125)TYSPALNKMF(134) (residues 24 to 33 of SEQ ID NO: 19)        binding stably to HLA molecules of the A24 type,    -   (129)ALNKMFCQL(137) (residues 28 to 36 of SEQ ID NO: 19) binding        stably to HLA molecules of the B35 type.

The invention also has for its object the polyepitopic fragment of thep53 human protein as defined above, characterized in that it correspondsto the fragment of 21 amino acids delimited by the amino acids locatedin the positions 149 and 169 of the peptide sequence of the p53 protein,and characterized by the following peptide sequence:

(149)STPPPGTRVRAMAIYKQSQHM(169) (SEQ ID NO: 20)

said fragment containing 6 epitopes binding stably to at least one ofthe 6 HLA molecules of the following types: A2, A3, A24, B27, B35 orB62, said epitopes being the following:

-   -   (149)STPPPGTRV(157) (residues 1 to 9 of SEQ ID NO: 20)binding        stably to HLA molecules of the A2 type,    -   (152)PPGTRVRAM(160) (residues 4 to 12 of SEQ ID NO: 20) binding        stably to HLA molecules of the B35 type,    -   (155)TRVRAMAIYK(164) (residues 7 to 16 of SEQ ID NO: 20) binding        stably to HLA molecules of the B27 type,    -   (156)RVRAMAIY(163) (residues 8 to 15 of SEQ ID NO: 20) binding        stably to HLA molecules of the B62 type,    -   (156)RVRAMAIYK(164) (residues 8 to 16 of SEQ ID NO: 20) binding        stably to HLA molecules of the A3 type,    -   (162)IYKQSQHM(169) (residues 14 to 21 of SEQ ID NO: 20) binding        stably to HLA molecules of the A24 type.

The invention also relates to the polyepitopic fragment of the p53 humanprotein as defined above, characterized in that it corresponds to thefragment of 26 amino acids delimited by the amino acids located inpositions 187 and 212 of the peptide sequence of the p53 protein, or tothe fragment of 34 amino acids delimited by the amino acids located inpositions 187 and 220 of said peptide sequence, this latter fragmentbeing characterized by the following peptide sequence:

(SEQ ID NO: 21) (187)GLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPY(220)

said fragment containing 11 epitopes binding stably to at least one ofthe 7 HLA molecules of the following types: A1, A2, A24, B7, B8, B27 orB44, said epitopes being the following:

-   -   (187)GLAPPQHLIRV(197) (residues 1 to 11 of SEQ ID NO: 21)        binding stably to HLA molecules of the A2 type,    -   (189)APPQHLIRV(197) (residues 3 to 11 of SEQ ID NO: 21) binding        stably to HLA molecules of the B7 type,    -   (195)IRVEGNLRVEY(205) (residues 9 to 19 of SEQ ID NO: 21)        binding stably to HLA molecules of the B27 type,    -   (196)RVEGNLRVEY(205) (residues 10 to 19 of SEQ ID NO: 21)        binding stably to HLA molecules of the A2 type,    -   (197)VEGNLRVEY(205) (residues 11 to 19 of SEQ ID NO: 21) binding        stably to HLA molecules of the B44 type,    -   (201)LRVEYLDDR(209) (residues 15 to 23 of SEQ ID NO: 21) binding        stably to HLA molecules of the B27 type,    -   (203)VEYLDDRNTF(212) (residues 17 to 26 of SEQ ID NO: 21)        binding stably to HLA molecules of the B44 type,    -   (204)EYLDDRNTF(212) (residues 18 to 26 of SEQ ID NO: 21) binding        stably to HLA molecules of the A24 type,    -   (210)NTFRHSVVV(218) (residues 24 to 32 of SEQ ID NO: 21) binding        stably to HLA molecules of the B8 type,    -   (21 1)TFRHSVVV(218) (residues 25 to 32 of SEQ ID NO: 21) binding        stably to HLA molecules of the A24 type,    -   (212)FRHSVVVPY(220) (residues 26 to 34 of SEQ ID NO: 21) binding        stably to HLA molecules of the B27 type.

The invention also has for its object the polyepitopic fragment of thep53 human protein as defined above, characterized in that it correspondsto the fragment of 18 amino acids delimited by the amino acids locatedin positions 226 and 243 of the peptide sequence of the p53 protein, andcharacterized by the following peptide sequence:

(226)GSDCTTIHYNYMCNSSCM(243) (SEQ ID NO: 22)

said fragment containing 3 epitopes binding stably to at least one ofthe 3 HLA molecules of the following types: A1, A24 or B44, saidepitopes being the following:

-   -   (226)GSDCTTIHY(234) (residues 1 to 9 of SEQ ID NO: 22) binding        stably to HLA molecules of the A1 type,    -   (227)SDCTTIHYNY(236) (residues 2 to 11 of SEQ ID NO: 22) binding        stably to HLA molecules of the B44 type,    -   (235)NYMCNSSCM(243) (residues 10 to 18 of SEQ ID NO: 22) binding        stably to HLA molecules of the A24 type.

The invention also relates to the polyepitopic fragment of the p53 humanprotein as defined above, characterized in that it corresponds to thefragment of 25 amino acids delimited by the amino acids located inpositions 249 and 273 of the peptide sequence of the p53 protein, or tothe fragment of 26 amino acids delimited by the amino acids located inpositions 248 and 273 of said peptide sequence, or to the fragment of 33amino acids delimited by the amino acids located in positions 248 and280 of said peptide sequence, this latter fragment being characterizedby the following peptide sequence:

(SEQ ID NO: 23) (248)RRPILTIITLEDSSGNLLGRNSFEVRVCACPGR(280)

said fragment containing 8 epitopes binding stably to at least one ofthe 6 HLA molecules of the following types: A2, B7, B27, B35, B44 orB62, said epitopes being the following:

-   -   (248)RRPILTIITL(257) (residues 1 to 10 of SEQ ID NO: 23) binding        stably to HLA molecules of the B27 type,    -   (249)RPILTIITL(257) (residues 2 to 10 of SEQ ID NO: 23) binding        stably to HLA molecules of the B35 and B7 type,    -   (255)ITLEDSSGN(263) (residues 8 to 16 of SEQ ID NO: 23) binding        stably to HLA molecules of the A2 type,    -   (257)LEDSSGNLL(265) (residues 10 to 18 of SEQ ID NO: 23) binding        stably to HLA molecules of the B44 type,    -   (263)NLLGRNSF(270) (residues 16 to 23 of SEQ ID NO: 23) binding        stably to HLA molecules of the B62 type,    -   (264)LLGRNSFEV(272) (residues 17 to 25 of SEQ ID NO: 23) binding        stably to HLA molecules of the A2 type,    -   (266)GRNSFEVR(273) (residues 19 to 26 of SEQ ID NO: 23) binding        stably to HLA molecules of the B27 type,    -   (272)VRVCACPGR(280) (residues 25 to 33 of SEQ ID NO: 23) binding        stably to HLA molecules of the B27 type.

The invention also relates to peptide sequences derived from thepolyepitopic fragments mentioned above, of the E6 or E7 proteins, or ofthe p53 protein, particularly:

-   -   by substitution and/or suppression and/or addition of one or        several amino acids, of the above-mentioned fragments, and/or    -   by modification of at least one peptide linkage —CO—NH— of the        peptide chain of the above-mentioned fragments, particularly by        introduction of a retro or retro-inverso type linkage, and/or    -   by substitution of at least one amino acid of the peptide chain        of the sequence or of the above-mentioned fragment, with a        non-proteinogenic amino acid,

said derived sequences containing peptides or pseudopeptides bindingspecifically to the same molecule or molecules of MCH as those bindingto the peptides contained in the polyepitopic fragments mentioned abovefrom which they derive.

By derived sequence by introduction of a retro-inverso linkage, shouldbe understood any peptide analog of an above-mentioned fragment, saidanalog being constituted by a peptide chain in which at least one of theresidues on the one hand is bound to at least one adjacent residue by an—NH—CO— linkage, and on the other hand, is of a chirality opposite thatof the same amino acyl residue in the peptide chain of the parentpeptide (namely of the above-mentioned fragment from which it derives).

By a sequence derived by introduction of a retro linkage, should beunderstood any peptide analog of an above-mentioned fragment, saidanalog being constituted by a peptide chain in which at least one of theresidues is bound to at least one adjacent residue by an —NH—CO—linkage, the chirality of the whole of the amino acyl residues involvedin at least one —NH—CO— linkage being conserved relative to thecorresponding residues of the peptide chain of the parent peptide.

It follows that the —CO—NH— and —NH—CO— linkages must be taken intoaccount in the preceding, in the direction of the parent peptide chaingoing from the amino terminal (N-terminal) end toward the carboxyterminal (C-terminal) end.

By “proteinogenic amino acid”, is meant, in the preceding, any aminoacid entering into the constitution of a natural protein or peptide.

By “non-proteinogenic amino acid” is meant, in contrast to the precedingdefinition, any amino acid that does not enter into the constitution ofa natural protein or peptide.

There is meant more particularly by “non-proteinogenic amino acid”, anyamino acid whose carbon carrying a R side chain, namely the —CHR— group,located between —CO— and —NH— in the natural peptide chain, is replacedby a structure that does not enter into the constitution of a naturalprotein or peptide.

The invention more particularly has for its object the derived sequencesas described above, characterized in that at least one of the peptidelinkages —CO—NH— of the peptide chain of the parent peptide is replacedby a linkage different from the —CO—NH— linkage, said different linkagebeing particularly selected from the following.

—CH₂—NH— (amino methylene);

—CH₂—CH₂— (carba);

—CO—CH₂— (cetomethylene);

—CH₂—O— (methylene-oxy);

—CHOH—CH₂— (hydroxyethylene);

—CHOH—CHOH— (di-hydroxyethylene);

—CH═CH— (E or Z olefin);

—CHCN—NH— (amino cyanomethylene);

—S—CH₂— (thiomethylene);

—CH₂—S— (thiomethylene);

—CS—NH— (thioamide);

—PO₂—NH— (phosphonamide);

—CHOH— (hydroxymethylene);

—NH—CO—NH— (urea);

(oxiran);

(tetrazole);

—CH₂—CO—NH— (β-homologation);

—CHOH—CH₂—NH— (amino hydroxyethylene);

—CO—NH—NH— (hydrazino).

The invention also has for its object nucleotide sequences coding for apolyepitopic fragment of the E6 or E7 protein, or for a derived peptidesequence, as defined above, said nucleotide sequences being derived fromthe sequence SEQ ID NO: 1 coding for the E6 protein, or from thesequence SEQ ID NO: 11 coding for the E7 protein.

In this connection, the invention more particularly has for its objectthe nucleotide sequences defined above, selected from the following:

-   -   the sequence SEQ ID NO: 3, coding for the polyepitopic fragment        SEQ ID NO: 4 mentioned above, of the E6 protein,    -   the sequence SEQ ID NO: 5, coding for the polyepitopic fragment        SEQ ID NO: 6 mentioned above, of the E6 protein,    -   the sequence SEQ ID NO: 7, coding for the polyepitopic fragment        SEQ ID NO: 8 mentioned above, of the E6 protein,    -   the sequence SEQ ID NO: 9, coding for the polyepitopic fragment        SEQ ID NO: 10 mentioned above, of the E6 protein,    -   the sequence SEQ ID NO: 13, coding for the polyepitopic fragment        SEQ ID NO: 14 mentioned above, of the E7 protein,    -   the sequence SEQ ID NO: 15, coding for the polyepitopic fragment        SEQ ID NO: 16 mentioned above, of the E7 protein,    -   the sequence SEQ ID NO: 17, coding for the polyepitopic fragment        SEQ ID NO: 18 mentioned above, of the E7 protein.

The invention also has for its object the nucleotide sequences codingfor a polyepitopic fragment of the p53 protein, or for a derived peptidesequence, as defined above.

The invention also has for its object any vector, particularly aplasmid, cosmid or phage, containing at least one above-mentionednucleotide sequence under the control of elements necessary for thetranscription of said sequence, particularly under the control of atranscription promoter and terminator.

The invention also relates to host cells, particularly bacterial, virus,yeasts, eucaryotic cells, transformed with the help of a vectormentioned above according to the invention, so as to integrate stablyinto their genome or to maintain in a stable manner in their cytoplasm,at least one nucleotide sequence according to the invention.

The invention also relates to any vector comprising one or severalpolyepitopic fragments and/or one or several derived peptide sequencesas defined above, or any vector comprising one or severalabove-mentioned nucleotide sequences, said vectors being selected fromthose adapted to ensure protection of said fragments or nucleotidesequences in the organism and/or their penetration into the cells of theorganism.

In the case of the use of polyepitopic fragments and/or theabove-mentioned derived peptide sequences, such vectors are selectedfrom fatty acids (in the framework of the preparation of lipopeptides),liposomes, etc.

In this connection, the invention more particularly has for its objectany lipopeptide characterized in that it comprises:

-   -   a peptide portion comprising one or several polyepitopic protein        fragments selected from those defined above, or any peptide        sequence derived from said fragments as defined above,    -   and one or several lipophilic portions, preferably selected from        those comprising:        -   a C4 to C20 hydrocarbon chain, saturated or unsaturated,            linear or branched,        -   or a steroid group, as the case may be connected to the            above-mentioned hydrocarbon chain,

said lipophilic portions being if desired associated with a shortpeptide vector (thereby to form lipopeptide vector structures)comprising one or several ionized functions at physiological pH, and afunction permitting the covalent bonding of said hydrocarbon chainand/or said steroid group.

By lipophilic portion, in what precedes and what follows, is intendedany lipophilic molecule, insoluble in water, permitting, when it islinked to the peptide portion defined above, an intracellular passivepassage of the obtained lipopeptide, thanks to the hydrophobicproperties of said molecule. Preferably the lipopeptide resulting fromthe linking of the lipophile portion to the peptide portion, is solublein water.

Preferably, the hydrocarbon chain of the lipophilic portions, isselected from the following:

-   -   palmitic acid,    -   oleic acid,    -   linoleic acid,    -   linolenic acid.

Also preferably, the steroid group of the lipophilic portion or portionsis selected from cholesterol derivatives such as cholest-5-enyl-3-oxyacetic acid, or cholest-5-enyl-3-oxycarbonic acid.

The invention more particularly has for its object any lipopeptide asdescribed above, characterized in that the lipophilic portion orportions are bonded covalently to one or several amino acids of thepeptide portion.

Preferably, the lypophilic portion or portions are bonded covalently tothe αNH₂ or εNH₂ function of a lysine located in the N terminal or Cterminal position of the peptide portion, or to the thiol function of acystein, or to any amino, alcohol or thiol function if desired added tothis peptide with a single spacer.

In this connection, the invention more particularly has for its objectany lipopeptide as defined above, in which the lipophilic portion orportions are represented by a group

N^(α)-acetyl-Lysine N^(ε)(palmitoyl) (also designated by theabbreviation Ac-K(Pam)).

The present invention also has for its object micelles ormicroaggregates of one or several different lipopeptides defined above.

Preferably, said micelles or microaggregates have a size less than about1 μm.

Preferably, the micelles or microaggregates according to the inventionare as obtained by dispersion of said lipopeptides in a concentratedacetic acid solution of about 80%, or any other solvent capable ofensuring molecular dispersion of the lipopeptides in solution.

In the case of the use of nucleotide sequences defined above accordingto the invention, the above-mentioned vectors are selected from theviruses, particularly the retroviruses, the adenoviruses and theassociated viruses (AAV Adeno Associated Virus).

The invention also has for its object antibodies directed against thepolyepitopic protein fragments or the epitopes or their derived peptidesequences (or analogs) as defined above, said antibodies being thoseobtained by immunization of an animal with at least one of theabove-mentioned complexes, said antibodies being adapted to form acomplex with these polyepitopic fragments or these epitopes or theiranalogs.

The antibodies according to the invention are polyclonal or monoclonalantibodies.

The polyclonal antibodies mentioned above are obtained by immunizationof an animal with at least one polyepitopic protein fragment or anepitope or an analog according to the invention, followed by therecovery of the desired antibodies in purified form, by removal of theserum of said animal, and separation of said antibodies from the otherconstituents of the serum, particularly by affinity chromatography on acolumn on which is fixed a specific antigen recognized by the antibody,particularly a polyepitopic protein fragment or an epitope or an analogaccording to the invention.

The monoclonal antibodies according to the invention can be obtained bythe hybridome technique whose general principle is set forth below.

In a first instance, an animal is immunized, generally a mouse (orculture cells in an in vitro immunization framework) with a polyepitopicprotein fragment or an epitope or an analog according to the invention,against which the B lymphocytes of the animal are then capable ofproducing antibodies. These antibody-producing lymphocytes are thenfused with “immortal” myelomatous cells (particularly of mice) to giverise to hybridomes. From the heterogeneous mixture of cells thusobtained, there is then carried out a selection of the cells capable ofproducing a particular antibody and of multiplying indefinitely. Thishybridome is multiplied in the form of clones, each leading to theproduction of a monoclonal antibody whose recognition propertiesrelative to the polyepitopic protein fragment or epitope or the like ofthe invention, can be tested for example with ELISA, by immunotransferin one or two dimensions, by immunofluorescence, or with the help of abiodetector. The monoclonal antibodies thus selected are then purifiedparticularly according to the affinity chromatography techniquedescribed above.

The invention also relates to the use of one or several above-mentionedantibodies for practicing a diagnostic method in vitro of theabove-mentioned pathologies.

In this connection, the invention also has for its object sets or kitscomprising said antibodies, for practicing a diagnostic method asdefined above.

The invention also relates to pharmaceutical compositions, or vaccines,characterized in that they comprise:

-   -   a)        -   at least one polyepitopic fragment of the E6 or E7 protein            as defined above,        -   and/or at least one peptide sequence derived from this            fragment, as defined above,        -   and/or at least one suitable vector, particularly            lipopeptides and/or micelles defined above, containing at            least one polyepitopic fragment mentioned above of the E6 or            E7 protein, and/or at least one sequence mentioned above            derived from these fragments,

in association with a physiologically acceptable vehicle,

said polyepitopic protein fragment and/or its derived sequence being, asthe case may be, associated with one or several other exogenous epitopesrecognized by auxiliary T cells (also called CD4 or T helper epitopes),said epitopes being selected particularly from the following:

-   -   the peptide fragment delimited by amino acids located in        positions 830 and 846 of the peptide sequence of the tetanus        toxin, said fragment responding to the following formula:        QYIKANSKFIGITELKK(SEQ ID NO: 24),    -   hemagglutinin (Prevost-Blondel et al., 1995, J. Virol, 62, No.        12, pp 8046-8055),    -   PADRE epitope (Alexander et al., 1994, Immunity, 1, 751).    -   or b)        -   at least one nucleotide sequence as defined above, coding            for an above-mentioned        -   polyepitopic fragment of the E6 or E7 protein,        -   and/or at least one nucleotide sequence coding for a peptide            sequence derived from this fragment, as defined above,

the above-mentioned nucleotide sequences being adapted to be used alone,as minigenes,

-   -   -   and/or at least one above-mentioned suitable vector,            selected particularly from the viruses such as defined            above, containing at least one above-mentioned nucleotide            sequence,

in association with a physiologically acceptable vehicle,

-   -   or c)        -   antibodies defined above, directed against a polyepitopic            fragment of the E6 or E7 protein, and/or against a peptide            sequence derived from these fragments, as defined above, in            association with a physiologically acceptable vehicle.

Preferably, the pharmaceutical compositions or vaccines mentioned aboveare present in a form administrable subcutaneously, particularly inseveral injections (preferably 3 injections) of about 500 μg of thepolyepitopic fragment in the lipopeptide form, at about one monthintervals.

The invention has more particularly for its object the use ofpolyepitopic fragments of the E6 or E7 protein defined above, or of theabove-mentioned derived peptide sequences, or the above-definednucleotide sequences, or the above-mentioned antibodies, or thelipopeptides defined above, for the preparation of a medication orvaccine for the prevention or treatment of pathologies connected withthe infection of individuals by human papillomavirus, such as cervicalintraepithelial neoplasias (CIN), the invasive cancer of the neck of theuterus, vulvar intraepithelial neoplasias (VIN).

The invention also relates to pharmaceutical compositions or vaccinescharacterized in that they comprise:

-   -   a)        -   at least one polyepitopic fragment of the p53 protein as            defined above,        -   and/or at least one peptide sequence derived from this            fragment, as defined above,        -   and/or at least one suitable vector, particularly the            lipopeptides and/or micelles defined above, containing at            least one above-mentioned polyepitopic fragment of the p53            protein, and/or at least one above-mentioned sequence            derived from these fragments,

in association with a physiologically acceptable vehicle,

said polyepitopic protein fragment and/or its derived sequence being, asthe case may be, associated with one or several other exogenous epitopesrecognized by the auxiliary T cells (also called CD4 or T helperepitopes), said epitopes being selected particularly from those definedabove,

-   -   or b)        -   at least one nucleotide sequence as defined above, coding            for an above-mentioned polyepitopic fragment of the p53            protein,        -   and/or at least one nucleotide sequence coding for a peptide            sequence derived from this fragment, as defined above,

the above-mentioned nucleotide sequences being adapted to be used alone,as minigenes,

-   -   -   and/or at least one above-mentioned suitable vector,            selected particularly from the viruses as defined above,            containing at least one above-mentioned nucleotide sequence,            in association with a physiologically acceptable vehicle,

    -   or c)        -   antibodies defined above, directed against a polyepitopic            fragment of the p53 protein, and/or against a peptide            sequence derived from these fragments, as defined above, in            association with a physiologically acceptable vehicle.

The invention also has for its object the use:

-   -   of at least one polyepitopic fragment of the p53 protein,        selected from those defined above,    -   and/or at least one peptide sequence derived from this fragment,        as defined above,    -   or at least one nucleotide sequence, as defined above, coding        for a polyepitopic fragment of the p53 protein, and/or for a        peptide sequence derived from this fragment, as defined above,

for the preparation of a medication or vaccine adapted for theprevention or treatment of cancers, particularly of the breast, of thecolon, of the lung or of the bladder.

The invention also relates to peptides or epitopes of the E6 protein ofHPV selected from the following:

-   -   (19)LPQLCTEL(26) (residues 19 to 26 of SEQ ID NO: 2) binding        stably to HLA molecules of the B51 type,    -   (21)QLCTELQTTI(30) (residues 21 to 30 of SEQ ID NO: 2) binding        stably to HLA molecules of the A2 type,    -   (24)TELQTTIHDI(33) (residues 24 to 33 of SEQ ID NO: 2) binding        stably to HLA molecules of the A29 or B44 type,    -   (33)IILECVYCK(41) (residues 33 to 41 of SEQ ID NO: 2) binding        stably to HLA molecules of the A11 type,    -   (35)LECVYCKQQL(44) (residues 35 to 44 of SEQ ID NO: 2) binding        stably to HLA molecules of the A29 or B44 type,    -   (37)CVYCKQQL(44) (residues 37 to 44 of SEQ ID NO: 2) binding        stably to HLA molecules of the B8 type,    -   (46)RREVYDFAFR(55) (residues 46 to 55 of SEQ ID NO: 2) binding        stably to HLA molecules of the B27 type,    -   (49)VYDFAFRDL(57) (residues 49 to 57 of SEQ ID NO: 2) binding        stably to HLA molecules of the A24 type,    -   (50)YDFAFRDL(57) (residues 50 to 57 of SEQ ID NO: 2) binding        stably to HLA molecules of the A29, B44 type,    -   (52)FAFRDLCIV(60) (residues 52 to 60 of SEQ ID NO: 2) binding        stably to HLA molecules of the A2, B35, B51 type,    -   (54)FRDLCIVYR(62) (residues 54 to 62 of SEQ ID NO: 2) binding        stably to HLA molecules of the A3, A11 type,    -   (59)IVYRDGNPY(67) (residues 59 to 67 of SEQ ID NO: 2) binding        stably to HLA molecules of the A3, A 11 type,    -   (81)SEYRHYCY(88) (residues 81 to 88 of SEQ ID NO: 2) binding        stably to HLA molecules of the A29, B44 type,    -   (87)CYSLYGTTL(95) (residues 87 to 95 of SEQ ID NO: 2) binding        stably to HLA molecules of the A24 type,    -   (94)TLEQQYNK(101) (residues 94 to 101 of SEQ ID NO: 2) binding        stably to HLA molecules of the A3, A11 type,    -   (95)LEQQYNKPL(103) (residues 95 to 103 of SEQ ID NO: 2) binding        stably to HLA molecules of the A29, B44 type,    -   (101)KPLCDLLI(108) (residues 101 to 108 of SEQ ID NO: 2) binding        stably to HLA molecules of the B7, B35, B51 type,    -   (118)CPEEKQRHL(126) (residues 118 to 126 of SEQ ID NO: 2)        binding stably to HLA molecules of the B8, B18, B35, B51 type,    -   (119)PEEKQRHL(126) (residues 119 to 126 of SEQ ID NO: 2) binding        stably to HLA molecules of the B44 type,    -   (127)DKKQRFHNI(135) (residues 127 to 135 of SEQ ID NO: 2)        binding stably to HLA molecules of the B8 type,    -   (128)KKQRFHNIR(136) (residues 128 to 136 of SEQ ID NO: 2)        binding stably to HLA molecules of the B27 type,    -   (130)QRFHNIRGRW(139) (residues 130 to 139 of SEQ ID NO: 2)        binding stably to HLA molecules of the B27 type,    -   (131)RFHNIRGRW(139) (residues 131 to 139 of SEQ ID NO: 2)        binding stably to HLA molecules of the A24 type.

The invention also relates to peptides or epitopes of the E7 protein ofHPV selected from the following:

-   -   (3)GDTPTLHEY(11) (residues 3 to 11 of SEQ ID NO: 12) binding        stably to HLA molecules of the B44 type,    -   (5)TPTLHEYML(13) (residues 5 to 13 of SEQ ID NO: 12) binding        stably to HLA molecules of the B35 type,    -   (15)LQPETTDLY(23) (residues 15 to 23 of SEQ ID NO: 12) binding        stably to HLA molecules of the B62 type,    -   (16)QPETTDLYCY(25) (residues 16 to 25 of SEQ ID NO: 12) binding        stably to HLA molecules of the A1, B18 type,    -   (45)AEPDRAHY(52) (residues 45 to 52 of SEQ ID NO: 12) binding        stably to HLA molecules of the A29, B44 type,    -   (46)EPDRAHYNIV(55) (residues 46 to 55 of SEQ ID NO: 12) binding        stably to HLA molecules of the B7 or B35 type,    -   (53)NIVTFCCK(60) (residues 53 to 60 of SEQ ID NO: 12) binding        stably to HLA molecules of the A3, A11 type,    -   (79)LEDLLMGTL(87) (residues 79 to 87 of SEQ ID NO: 12) binding        stably to HLA molecules of the A29, B44 type,    -   (89)IVCPICSQK(97) (residues 89 to 97 of SEQ ID NO: 12) binding        stably to HLA molecules of the A3, A11 type.

The invention also relates to peptide sequences derived from theabove-mentioned peptides, said derived sequences, or the like, being asdefined above in the framework of sequences derived from thepolyepitopic protein fragments mentioned above.

The invention also has for its object the nucleotide sequences codingfor peptides of the E6 or E7 proteins mentioned above, namely,

-   -   the sequence delimited by the nucleotides located in positions        43 and 66 of the sequence SEQ ID NO: 1, coding for        (15)RPRKLPQL(22) (residues 15 to 22 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        52 and 78 of the sequence SEQ ID NO: 1, coding for        (18)KLPQLCTEL(26) (residues 18 to 26 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        55 and 78 of the sequence SEQ ID NO: 1, coding for        (19)LPQLCTEL(26) (residues 19 to 26 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        61 and 90 of the sequence SEQ ID NO: 1, coding for        (21)QLCTELQTTI(30) (residues 21 to 30 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        70 and 99 of the sequence SEQ ID NO: 1, coding for        (24)TELQTTIHD(33) (residues 24 to 33 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        97 and 123 of the sequence SEQ ID NO: 1, coding for        (33)IILECVYCK(41) (residues 33 to 41 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        103 and 132 of the sequence SEQ ID NO: 1, coding for        (35)LECVYCKQQL(44) (residues 35 to 44 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        109 and 132 of the sequence SEQ ID NO: 1, coding for        (37)CVYCKQQL(44) (residues 37 to 44 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        136 and 165 of the sequence SEQ ID NO: 1, coding for        (46)RREVYDFAFR(55) (residues 46 to 55 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        145 and 171 of the sequence SEQ ID NO: 1, coding for        (49)VFDFAFRDL(57) (residues 49 to 57 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        148 and 171 of the sequence SEQ ID NO: 1, coding for        (50)YDFAFRDL(57) (residues 50 to 57 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        154 and 180 of the sequence SEQ ID NO: 1, coding for        (52)FAFRDLCIV(60) (residues 52 to 60 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        160 and 186 of the sequence SEQ ID NO: 1, coding for        (54)FRDLCIVYR(62) (residues 54 to 62 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        175 and 201 of the sequence SEQ ID NO: 1, coding for        (59)IVYRDGNPY(67) (residues 59 to 67 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        241 and 264 of the sequence SEQ ID NO: 1, coding for        (81)SEYRHYCY(88) (residues 81 to 88 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        259 and 285 of the sequence SEQ ID NO: 1, coding for        (87)CYRLYGTTL(95) (residues 87 to 95 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        280 and 303 of the sequence SEQ ID NO: 1, coding for        (94)TLEQQYNK( 101) (residues 94 to 101 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        283 and 309 of the sequence SEQ ID NO: 1, coding for        (95)LEQQYNKPL(103) (residues 95 to 103 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        301 and 324 of the sequence SEQ ID NO: 1, coding for        (101)KPLCDLLI(108) (residues 101 to 108 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        352 and 378 of the sequence SEQ ID NO: 1, coding for        (118)CPEEKQRHL(126) (residues 118 to 126 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        355 and 378 of the sequence SEQ ID NO: 1, coding for (        19)PEEKQRHL(126) (residues 119 to 126 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        379 and 405 of the sequence SEQ ID NO: 1, coding for        (127)DKKQRFHNI(135) (residues 127 to 135 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        382 and 408 of the sequence SEQ ID NO: 1, coding for        (128)KKQRFHNIR(136) (residues 128 to 136 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        388 and 417 of the sequence SEQ ID NO: 1, coding for        (130)QRFHNIRGRW(139) (residues 130 to 139 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions        391 and 417 of the sequence SEQ ID NO: 1, coding for        (131)RFHNIRGRW(139) (residues 131 to 139 of SEQ ID NO: 2),    -   the sequence delimited by the nucleotides located in positions 7        and 33 of the sequence SEQ ID NO: 11, coding for        (3)GDTPTLHEY(11) (residues 3 to 11 of SEQ ID NO: 12),    -   the sequence delimited by the nucleotides located in positions        13 and 39 of the sequence SEQ ID NO: 11, coding for        (5)TPTLHEYML(13) (residues 5 to 13 of SEQ ID NO: 12),    -   the sequence delimited by the nucleotides located in positions        43 and 69 of the sequence SEQ ID NO: 11, coding for        (15)LQPETTDLY(23) (residues 15 to 23 of SEQ ID NO: 12),    -   the sequence delimited by the nucleotides located in positions        46 and 75 of the sequence SEQ ID NO: 11, coding for        (16)QPETTDLYCY(25) (residues 16 to 25 of SEQ ID NO: 12),    -   the sequence delimited by the nucleotides located in positions        133 and 153 of the sequence SEQ ID NO: 11, coding for        (45)AEPDRAHY(52) (residues 45 to 52 of SEQ ID NO: 12),    -   the sequence delimited by the nucleotides located in positions        136 and 165 of the sequence SEQ ID NO: 11, coding for        (46)EPDRAHYNIV(55) (residues 46 to 55 of SEQ ID NO: 12),    -   the sequence delimited by the nucleotides located in positions        157 and 180 of the sequence SEQ ID NO: 11, coding for        (53)NIVTFCCK(60) (residues 53 to 60 of SEQ ID NO: 12),    -   the sequence delimited by the nucleotides located in positions        235 and 261 of the sequence SEQ ID NO: 11, coding for        (79)LEDLLMGTL(87) (residues 79 to 87 of SEQ ID NO: 12),    -   the sequence delimited by the nucleotides located in positions        265 and 291 of the sequence SEQ ID NO: 11, coding for        (89)IVCPICSQK(97) (residues 89 to 97 of SEQ ID NO: 12).

The invention also has for its object epitopes of the p53 proteinselected from the following:

-   -   (102)TYQGSYGFRL(111) (residues 1 to 10 of SEQ ID NO: 19) binding        stably to HLA molecules of the A24 type,    -   (105)GSYGFRLGFL(114) (residues 4 to 13 of SEQ ID NO: 19) binding        stably to HLA molecules of the B35 type,    -   (106)SYGFRLGFL(114) (residues 5 to 13 of SEQ ID NO: 19) binding        stably to HLA molecules of the A24 type,    -   (118)TAKSVTCTY(126) (residues 17 to 25 of SEQ ID NO: 19) binding        stably to HLA molecules of the B62 type,    -   (125)TYSPALNKMF(134) (residues 24 to 33 of SEQ ID NO: 19)        binding stably to HLA molecules of the A24 type,    -   (152)PPGTRVRAM(160) (residues 4 to 12 of SEQ ID NO: 20) binding        stably to HLA molecules of the B35 type,    -   (155)TRVRAMAIYK(164) (residues 7 to 16 of SEQ ID NO: 20) binding        stably to HLA molecules of the B27 type,    -   (156)RVRAMAIY(163) (residues 8 to 15 of SEQ ID NO: 20) binding        stably to HLA molecules of the B62 type,    -   (162)IYKQSQHM(169) (residues 14 to 21 of SEQ ID NO: 20) binding        stably to HLA molecules of the A24 type,    -   (195)IRVEGNLRVEY(205) (residues 9 to 19 of SEQ ID NO: 21)        binding stably to HLA molecules of the B27 type,    -   (197)VEGNLRVEY(205) (residues 11 to 19 of SEQ ID NO: 21) binding        stably to HLA molecules of the B44 type,    -   (201)LRVEYLDDR(209) (residues 15 to 23 of SEQ ID NO: 21) binding        stably to HLA molecules of the B27 type,    -   (203)VEYLDDRNTF(212) (residues 17 to 26 of SEQ ID NO: 21)        binding stably to HLA molecules of the B44 type,    -   (204)EYLDDRNTF(212) (residues 18 to 26 of SEQ ID NO: 21) binding        stably to HLA molecules of the A24 type,    -   (211)TFRHSVVV(218) (residues 25 to 32 of SEQ ID NO: 21) binding        stably to HLA molecules of the A24 type,    -   (212)FRHSVVVPY(220) (residues 26 to 34 of SEQ ID NO: 21) binding        stably to HLA molecules of the B27 type,    -   (227)SDCTTIHYNY(236) (residues 2 to 11 of SEQ ID NO: 22) binding        stably to HLA molecules of the B44 type,    -   (235)NYMCNSSCM(243) (residues 10 to 18 of SEQ ID NO: 22) binding        stably to HLA molecules of the A24 type,    -   (249)RPILTIITL(257) (residues 2 to 10 of SEQ ID NO: 23) binding        stably to HLA molecules of the B35 type,    -   (257)LEDSSGNLL(265) (residues 10 to 18 of SEQ ID NO: 23) binding        stably to HLA molecules of the B44 type,    -   (263)NLLGRNSF(270) (residues 16 to 23 of SEQ ID NO: 23) binding        stably to HLA molecules of the B62 type,    -   (266)GRNSFEVR(273) (residues 19 to 26 of SEQ ID NO: 23) binding        stably to HLA molecules of the B27 type,    -   (272)VRVCACPGR(280) (residues 25 to 33 of SEQ ID NO: 23) binding        stably to HLA molecules of the B27 type.

The invention also has for its object any process for the preparation ofpolyepitopic fragments, of single epitopes (above-mentioned peptides),or of derived sequences, by conventional peptide synthesis in liquid orsolid phase.

As a modification, the polyepitopic fragments, single epitopes orderived peptide sequences, as defined above according to the invention,can be obtained in the form of recombinant polypeptides bytransformation of suitable host cells as defined above with the help ofvectors containing a recombinant nucleotide sequence as defined aboveaccording to the invention, and the recovery, as the case may be afterpurification, of the recombinant polypeptide coded by said nucleotidesequence and produced by the host cells mentioned above.

1. Polyepitopic fragments of the E6 or E7 protein of HPV, characterizedin that they comprise a peptide sequence of about 15 to 30 amino acids,this peptide sequence containing amino acid sequences of at least 3different epitopes binding stably to HLA molecules of identical ordifferent type, when these epitopes are obtained by enzymaticdegradation of said peptide sequence, particularly in the proteasome,such that at least 4 HLA molecules of different types bind to theseepitopes, these 4 HLA molecules being selected from among those of typesA1, A2, A3, A11, A24, A29, B7, B8, B18, B27, B35, B44, B51 and B62. 2.Polyepitopic fragments according to claim 1, characterized in that thenumber of amino acids of their peptide sequence is greater than or equalto 17, and less than or equal to
 30. 3. Polyepitopic fragments of the E6protein of HPV according to claim 1, characterized in that they comprisea peptide sequence of about 15 to 30 amino acids, this peptide sequencecontaining amino acid sequences of at least 5 different epitopes bindingstably to HLA molecules of identical or different type, when theseepitopes are obtained by enzymatic degradation of said peptide sequence,particularly in the proteasome, such that at least 6 HLA molecules ofdifferent types bind to these epitopes, these 6 HLA molecules beingselected from those of types A1, A2, A3, A11, A24, A29, B7, B8, B18,B27, B35, B44 and B51.
 4. Polyepitopic fragments of the E6 protein ofHPV according to one of claim 1, characterized in that they all comprisean epitope binding to the HLA molecule of type B35, an epitope bindingto the HLA molecule of type B44, and an epitope binding to the HLAmolecule of type B51.
 5. Polyepitopic fragment of the E6 protein of HPVaccording to claim 1, characterized by the following: (15)RPRKLPQL(22)(residues 15 to 22 of SEQ ID NO: 2) binding stably to HLA molecules ofthe B7 or B35 type, (18)KLPQLCTEL(26) (residues 18 to 26 of SEQ ID NO:2) binding stably to HLA molecules of the A2 type, (19)LPQLCTEL(26)(residues 19 to 26 of SEQ ID NO: 2) binding stably to HLA molecules ofthe B51 type, (21)QLCTELQTTI(30) (residues 21 to 30 of SEQ ID NO: 2)binding stably to HLA molecules of the A2 type, (24)TELQTTIHDI(33)(residues 24 to 33 of SEQ ID NO: 2) binding stably to HLA molecules ofthe A29 or B44 type, (29)TIHDIILRCV(38) (residues 29 to 38 of SEQ ID NO:2) binding stably to HLA molecules of the A2 type, (33)IILECVYCK(41)(residues 33 to 41 of SEQ ID NO: 2) binding stably to HLA molecules ofthe A11 type, (35)LECVYCKQQL(44) (residues 35 to 44 of SEQ ID NO: 2)binding stably to HLA molecules of the A29 or B44 type, (37)CVYCKQQL(44)(residues 37 to 44 of SEQ ID NO: 2) binding stably to HLA molecules ofthe B8 type.
 6. A polyepitopic fragment comprising SEQ ID NO: 8 or SEQID NO:
 10. 7. Polyepitopic fragment of the E6 protein of HPV accordingto claim 1, characterized in that it corresponds to the fragment of 29amino acids delimited by the amino acids located in positions 80 and 108of the peptide sequence of the E6 protein of HPV, this latter fragmentbeing characterized by the peptide sequence SEQ ID NO: 8 as follows:(80)ISEYRHYCYSLYGTTLEQQYNKPLCDLLI(108)

said fragment containing 6 epitopes binding stably to at least one ofthe 10 HLA molecules of the following types: A1, A3, A11, A24, A29, B7,B18, B35, B44, or B51, said epitopes being the following:(80)ISEYRHYCY(88) (residues 80 to 88 of SEQ ID NO: 2) binding stably toHLA molecules of the A1 or B18 type, (81)SEYRHYCY(88) (residues 81 to 88of SEQ ID NO: 2) binding stably to HLA molecules of the A29 or B44 type,(87)CYSLYGTTL(95) (residues 87 to 95 of SEQ ID NO: 2) binding stably toHLA molecules of the A24 type, (94)TLEQQYNK(101) (residues 94 to 101 ofSEQ ID NO: 2) binding stably to HLA molecules of the A3 or A11 type,(95)LEQQYNKPL(103) (residues 95 to 103 of SEQ ID NO: 2) binding stablyto HLA molecules of the A29 or B44 type, (11)KPLCDLLI(108) (residues 101to 108 of SEQ ID NO: 2) binding stably to HLA molecules of the B7, B35or B51 type.
 8. Polyepitopic fragment of the E6 protein of HPV accordingto claim 1, characterized in that it corresponds to the fragment of 22amino acids delimited by the amino acids located in positions 118 and139 of the peptide sequence of the E6 protein of HPV, this latterfragment being characterized by the peptide sequence SEQ ID NO: 10 asfollows: (118)CPEEKQRHLDKKQRFHNIRGRW(139)

said fragment containing 6 epitopes binding stably to at least one ofthe 7 HLA molecules of the following types: A24, B8, B18, B27, B35, B44,or B51, said epitopes being the following: (118)CPEEKQRHL(126) (residues118 to 126 of SEQ ID NO: 2) binding stably to HLA molecules of the B8,B18, B35, B51 type, (119)PEEKQRHL(126) (residues 119 to 126 of SEQ IDNO: 2) binding stably to HLA molecules of the B44 type,(127)DKKQRFHNI(135) (residues 127 to 135 of SEQ ID NO: 2) binding stablyto HLA molecules of the B8 type, (128)KKQRFHNIR(136) (residues 128 to136 of SEQ ID NO: 2) binding stably to HLA molecules of the B27 type,(130)QRFHNIRGRW(139) (residues 130 to 139 of SEQ ID NO: 2) bindingstably to HLA molecules of the B27 type, (131)RFHNIRGRW(139) (residues131 to 139 of SEQ ID NO: 2) binding stably to HLA molecules of the A24type.
 9. Polyepitopic fragments of the E7 protein of HPV according toclaim 1, characterized in that they comprise a peptide sequence of about15 to 30 amino acids, this peptide sequence containing amino acidsequences of at least 3 different epitopes binding stably to HLAmolecules of identical or different type, when these epitopes areobtained by enzymatic degradation of said peptide sequence, particularlyin the proteasome, such that at least 4 HLA molecules of different typesbind to these epitopes, these 4 HLA molecules being selected from thoseof type A1, A2, A3, A11, A29, E7, B18, B35, B44 and B62. 10.Polyepitopic fragments of the E7 protein of HPV according to claim 9,characterized in that they all comprise an epitope binding to the HLAmolecule of type B44.
 11. Polyepitopic fragment of the E7 protein of HPVaccording to claim 9, characterized by the following: (3)GDTPTLHEY(11)(residues 3 to 11 of SEQ ID NO: 12) binding stably to HLA molecules ofthe B44 type, (5)TPTLHEYML(13) (residues 5 to 13 of SEQ ID NO: 12)binding stably to HLA molecules of the B35 type, (11)YMLDLQPETT(20)(residues 11 to 20 of SEQ ID NO: 12) binding stably to HLA molecules ofthe A2 type, (15) LQPETTDLY(23) (residues 15 to 23 of SEQ ID NO: 12)binding stably to HLA molecules of the B62 type, (16)QPETTDLYCY(25)(residues 16 to 25 of SEQ ID NO: 12) binding stably to HLA molecules ofthe A1 or B18 type.
 12. Polyepitopic fragment of the E7 protein of HPVaccording to claim 9, characterized in that it corresponds to thefragment of 17 amino acids delimited by the amino acids located inpositions 44 and 60 of the peptide sequence of the E7 protein of HPV,this latter fragment being characterized by the peptide sequence SEQ IDNO: 16 as follows: (44)QAEPDRAHYNIVTFCCK(60)

said fragment containing 4 epitopes binding stably to at least one ofthe 6 HLA molecules of the following types: A1, A3, A11, A29, B7, B18,B35, or B44, said epitopes being the following: (44)QAEPDRAHY(52)(residues 44 to 52 of SEQ ID NO: 12) binding stably to HLA molecules ofthe A1 or B18 type, (45)AEPDRAHY(52) (residues 45 to 52 of SEQ ID NO:12) binding stably to HLA molecules of the A29 or B44 type,(46)EPDRAHYNIV(55) (residues 46 to 55 of SEQ ID NO: 12) binding stablyto HLA molecules of the B7 or B35 type, (53)NIVTFCCK(60) (residues 53 to60 of SEQ ID NO: 12) binding stably to HLA molecules of the A3 or A11type.
 13. Polyepitopic fragment of the E7 protein of HPV according toclaim 9, characterized in that it corresponds to the fragment of 19amino acids delimited by the amino acids located in positions 79 and 97of the peptide sequence of the E7 protein of HPV, this latter fragmentbeing characterized by the peptide sequence SEQ ID NO: 18 as follows:(79)LEDLLMGTLGIVCPICSQK(97)

said fragment containing 4 epitopes binding stably to at least one ofthe 5 HLA molecules of the following types: A2, A3, A11, A29 or B44,said epitopes being the following: (79)LEDLLMGTL(87) (residues 79 to 87of SEQ ID NO: 12) binding stably to HLA molecules of the A29 or B44type, (82) LLMGTLGIV(90) (residues 82 to 90 of SEQ ID NO: 12) bindingstably to HLA molecules of the A2 type, (86)TLGIVCPI(93) (residues 86 to93 of SEQ ID NO: 12) binding stably to HLA molecules of the A2 type,(89) IVCPICSQK(97) (residues 89 to 97 of SEQ ID NO: 12) binding stablyto HLA molecules of the A3 or A11 type.
 14. Polyepitopic fragments ofthe E6 or E7 protein, characterized in that they correspond to thepeptide sequences derived from the polyepitopic fragments according toclaim 1, by substitution, and/or suppression, and/or addition of one orseveral amino acids, of the above-mentioned fragments, and/or bymodification of at least one —CO—NH— peptide linkage of the peptidechain of the above-mentioned fragments, particularly by introduction ofa retro or retro-inverso type linkage, and/or by substitution of atleast one amino acid of the peptide chain of the sequence or of theabove-mentioned fragment, with a non-proteinogenic amino acid, saidderived sequences containing peptides or pseudopeptides bindingspecifically to the same molecule or molecules of MCH as those bindingto the peptides contained in the above-mentioned polyepitopic fragmentsfrom which they derive.
 15. Nucleotide sequences coding for apolyepitopic fragment or for a peptide sequence derived according toclaim 1, said nucleotide sequences being derived from SEQ ID NO: 1coding for the E6 protein, or from SEQ ID NO: 11 coding for the E7protein.
 16. Nucleotide sequences according to claim 15, selected fromthe following: the sequence SEQ ID NO: 5, coding for the polyepitopicfragment SEQ ID NO: 6, the sequence SEQ ID NO: 7, coding for thepolyepitopic fragment SEQ ID NO: 8, the sequence SEQ ID NO: 9, codingfor the polyepitopic fragment SEQ ID NO: 10, the sequence SEQ ID NO: 15,coding for the polyepitopic fragment SEQ ID NO: 16, the sequence SEQ IDNO: 17, coding for the polyepitopic fragment SEQ ID NO:
 18. 17.Polyclonal or monoclonal antibodies, directed against a polyepitopicfragment or against a peptide sequence derived according to claim
 1. 18.Lipopeptide characterized in that said lipopeptide comprises: a peptideportion comprising one or several polyepitopic protein fragments, or apeptide sequence derived from said fragments, as defined in one ofclaims 1, and one or several lipophile portions, such as thosecomprising: a C4 to C20 hydrocarbon chain, saturated or unsaturated,linear or branched, or a steroid group, as the case may be bonded to theabove-mentioned hydrocarbon chain, said lipophilic portions being ifdesired associated with a short peptide vector comprising one or severalionized functions at physiological pH, and a function permitting thecovalent bonding of said hydrocarbon chain and/or said steroid group.19. A pharmaceutical composition, or vaccine, characterized in that itcomprises: at least one polyepitopic fragment of the E6 or E7 accordingto claim 1, and/or at least one peptide sequence derived from saidfragment, and/or at least one suitable vector, containing saidpolyepitopic fragment of the E6 or E7 protein, and/or at least one ofsaid sequence derived from these fragments, in association with aphysiologically acceptable vehicle, said polyepitopic protein fragmentand/or its derived sequence being, as the case may be, associated withone or several other exogenous epitopes recognized by auxiliary T cells,such as the peptide fragment delimited by the amino acids located inpositions 830 and 846 of the peptide sequence of the tetanus toxin,hemagglutinin, or PADRE epitope.
 20. A pharmaceutical composition, orvaccine, characterized in that said composition comprises: at least onenucleotide sequence, coding for polyepitopic fragment of the E6 or E7protein, and/or at least one nucleotide sequence coding for a peptidesequence derived from said fragment, and/or at least one suitablevector, in association with a physiologically acceptable vehicle.
 21. Apharmaceutical composition, or vaccine, characterized in that itcomprises: antibodies according to claim 17, directed against apolyepitopic fragment of the E6 or E7 protein, and/or against a peptidesequence derived from these fragments, as defined above.
 22. Epitopes ofthe E6 protein of HPV selected from the following: (19)LPQLCTEL(26)(residues 19 to 26 of SEQ ID NO: 2) binding stably to HLA molecules ofthe B51 type, (21)QLCTELQTTI(30) (residues 21 to 30 of SEQ ID NO: 2)binding stably to HLA molecules of the A2 type, (24)TELQTTIHDI(33)(residues 24 to 33 of SEQ ID NO: 2) binding stably to HLA molecules ofthe A29 or B44 type, (33)IILECVYCK(41) (residues 33 to 41 of SEQ ID NO:2) binding stably to HLA molecules of the A11 type, (35)LECVYCKQQL(44)(residues 35 to 44 of SEQ ID NO: 2) binding stably to HLA molecules ofthe A29 or B44 type, (37)CVYCKQQL(44) (residues 37 to 44 of SEQ ID NO:2) binding stably to HLA molecules of the B8 type, (46)RREVYDFAFR(55)(residues 46 to 55 of SEQ ID NO: 2) binding stably to HLA molecules ofthe B27 type, (49)VYDFAFRDL(57) (residues 49 to 57 of SEQ ID NO: 2)binding stably to HLA molecules of the A24 type, (50)YDFAFRDL(57)(residues 50 to 57 of SEQ ID NO: 2) binding stably to HLA molecules ofthe A29, B44 type, (52)FAFRDLCIV(60) (residues 52 to 60 of SEQ ID NO: 2)binding stably to HLA molecules of the A2, B35, B51 type,(54)FRDLCIVYR(62) (residues 54 to 62 of SEQ ID NO: 2) binding stably toHLA molecules of the A3, A11 type, (59)IVYRDGNPY(67) (residues 59 to 67of SEQ ID NO: 2) binding stably to HLA molecules of the A3, A11 type,(81)SEYRHYCY(88) (residues 81 to 88 of SEQ ID NO: 2) binding stably toHLA molecules of the A29, B44 type, (87)CYSLYGTTL(95) (residues 87 to 95of SEQ ID NO: 2) binding stably to HLA molecules of the A24 type,(94)TLEQQYNK(101) (residues 94 to 101 of SEQ ID NO: 2) binding stably toHLA molecules of the A3, A11 type, (95)LEQQYNKPL(103) (residues 95 to103 of SEQ ID NO: 2) binding stably to HLA molecules of the A29, B44type, (101)KPLCDLLI(108) (residues 101 to 108 of SEQ ID NO: 2) bindingstably to HLA molecules of the B7, B35, B51 type, (118)CPEEKQRHL(126)(residues 118 to 126 of SEQ ID NO: 2) binding stably to HLA molecules ofthe B8, B18, B35, B51 type, (119)PEEKQRHL(126) (residues 119 to 126 ofSEQ ID NO: 2) binding stably to HLA molecules of the B44 type,(127)DKKQRFHNI(135) (residues 127 to 135 of SEQ ID NO: 2) binding stablyto HLA molecules of the B8 type, (128)KKQRFHNIR(136) (residues 128 to136 of SEQ ID NO: 2) binding stably to HLA molecules of the B27 type,(130)QRFHNIRGRW(139) (residues 130 to 139 of SEQ ID NO: 2) bindingstably to HLA molecules of the B27 type, (131)RFHNIRGRW(139) (residues131 to 139 of SEQ ID NO: 2) binding stably to HLA molecules of the A24type.
 23. Epitopes of the E7 protein of HPV selected from the following:(3)GDTPTLHEY(11) (residues 3 to 11 of SEQ ID NO: 12) binding stably toHLA molecules of the B44 type, (5)TPTLHEYML(13) (residues 5 to 13 of SEQID NO: 12) binding stably to HLA molecules of the B35 type,(15)LQPETTDLY(23) (residues 15 to 23 of SEQ ID NO: 12) binding stably toHLA molecules of the B62 type, (16)QPETTDLYCY(25) (residues 16 to 25 ofSEQ ID NO: 12) binding stably to HLA molecules of the A1, B18 type,(45)AEPDRAHY(52) (residues 45 to 52 of SEQ ID NO: 12) binding stably toHLA molecules of the A29, B44 type, (46)EPDRAHYNIV(55) (residues 46 to55 of SEQ ID NO: 12) binding stably to HLA molecules of the B7 or B35type, (53)NIVTFCCK(60) (residues 53 to 60 of SEQ ID NO: 12) bindingstably to HLA molecules of the A3, A11 type, (79)LEDLLMGTL(87) (residues79 to 87 of SEQ ID NO: 12) binding stably to HLA molecules of the A29,B44 type, (89)IVCPICSQK(97) (residues 89 to 97 of SEQ ID NO: 12) bindingstably to HLA molecules of the A3, A11 type.